Perinatal brain injury resulting in neurodevelopmental delay (ND), cerebral palsy (CP) and seizures represents the one of most severe disabilities in childhood. The incidence of CP is 40-148 in preterm and 1-2/1,000 in full term infants. Developmental disabilities place a huge burden on society (lifetime costs per person: ~1 million dollars), emphasizing the urgent need for improved treatment strategies to reduce perinatal brain damage. Unfortunately, hypothermia is the only approved therapy for brain damage in infants and is only partially protective. It can only be used to treat hypoxic-ischemic (HI) encephalopathy in full term infants, and cannot be used in preterm infants in whom supportive care is the only `therapy' to attenuate brain damage. Cytokines represent a final common pathway, which cause/augment fetal/neonatal brain damage. Inter-alpha inhibitor proteins (IAIPs) down-regulate pro-inflammatory cytokines in sepsis and inhibit destructive serine proteases. Little information is available about IAIPs in brain. Recent data suggests that bikunin, a fragment of IAIPs, isolated from urine, attenuates stroke-related brain injury and experimental autoimmune encephalomyelitis-related white matter loss in adult rats. However, the half-life of bikunin is very short (3-10 min), compared with the complexed form isolated from blood (18 h in neonatal rats), requiring large quantities of protein and continuous intravenous infusions. The potential neuroprotective effects of IAIPs isolated from blood have not been examined except for our recent data. We will examine the neuroprotective effects of the blood-derived IAIPs. We anticipate that this form will be a more viable neuroprotective agent for clinical use. Our preliminary studies with blood-derived IAIPs suggest that this agent has remarkable neuroprotective effects in HI neonatal rats and fetal sheep after ischemic injury. Our overall goal is to develop a novel effective therapy to treat ischemic brain damage using the preclinical fetal sheep model in which IAIPs can be given by the clinically relevant intravenous route. We hypothesize that systemic IAIPs administration attenuates the development of ischemic-reperfusion related injury in the immature brain. The purpose of this proposal is to provide a strong biological basis to support the use of IAIPs as therapeutic agents to treat ischemia-related brain injury in the immature brain. The aims are: (1) To establish the dose of IAIPs with the greatest neuroprotective efficacy in the fetus; (2) To examine the neuroprotective efficacy of delayed treatment with IAIPs on brain injury; (3) To determine the pharmacokinetics of intravenously administered IAIPs in the ovine fetus. Fetuses will be surgically prepared; brain ischemia induced by carotid occlusion and injury measured by a multidisciplinary approach using ECoG, physiological, biochemical, pathological, immunological, immunohistochemical, and molecular methods. Results of the studies could yield novel information that will accelerate the use of IAIPs as neuroprotective agents to treat brain injury in the fetus/neonate potentially t prevent ND and CP in infants.